MILLIPLEX® MAP Frequently Asked Questions

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1. Do you have a resource guide for using MILLIPLEX® MAP assay panels?

For a comprehensive list of our MILLIPLEX® MAP panels and kits, download our Analyte Quarterly and our most comprehensive Tips and Tricks brochure.

For additional assistance, contact our Technical Support Team.

2. What do you mean by analyte?

In our assays, an “analyte” is a specific protein biomarker of interest which you can quantify using our MILLIPLEX® MAP assay kits.

3. What is MILLIPLEX® MAP and how does the technology work?

MILLIPLEX® MAP uses the proprietary Luminex® xMAP® bead-based multiplex assay platform. For example, imagine if the enzyme-linked immunosorbent assay (ELISA or EIA) was on a bead, instead of a plate. Each magnetic MagPlex® microsphere bead is fluorescently coded with one of 80 specific ratios of two fluorophores (each spectrally distinct set is known as a bead region, fluorescent at ex 635 nm). Analyte-specific capture antibodies are bound to beads of a specific region, then unbound sites of the bead are blocked. Beads are incubated with sample, washed and then incubated with a biotinylated secondary antibody. The unbound secondary antibody is washed away and the added streptavidin-phycoerythrin binds to the biotin of the “sandwich” immunoassay. The Luminex® instrument detects individual beads by region plus the streptavidin-conjugated R-Phycoerythrin (SAPE) signal, ex 525 nm, indicating the analyte is present. Using this ELISA on a bead method, in theory, you could detect and quantify up to 80 different analytes in one small sample using the Luminex® 200™ or FLEXMAP 3D® instruments, or up to 50 analytes using the MAGPIX® instrument.

4. How do the detection systems work?

There are two different detection systems used by Luminex®. The Luminex® 200™ and FLEXMAP 3D® instruments use a flow cytometry-based system where beads flow in sheath fluid through a cell with a red laser (635 nm) identifying the bead region and a green laser (525 nm) quantifying the bound analyte “sandwich.” The MAGPIX® instrument uses a CCD imager to identify beads and quantify analytes with red LED (635 nm) and green LED (525 nm) light sources.

5. Do I have to use a Luminex® instrument?

A Luminex® instrument is required to run Luminex®-based immunoassays, including MILLIPLEX® MAP assays and kits. These instruments may be obtained from multiple Luminex® partners. Note that the Bio-Plex® instrument may also be used to run any Luminex®-based immunoassay.

6. Are your MILLIPLEX® MAP assay kits validated?

Our MILLIPLEX® MAP assay kits are the most highly validated research use only (RUO) multiplex kits available on the market. We provide all assay performance data with every protocol: sensitivity, specificity, linearity, precision, accuracy, and stability. With every kit we provide an optimized serum matrix to add to each standard well so that every sample interrogated is compared to standards in a sample-matched matrix. Our lot-to-lot reproducibility is matched back to a “Gold Standard” lot. Our trend charts and lot records are available upon request.

Our kits are for research use only. Not for use in diagnostic procedures.

7. How many analytes and detection kits do you offer?

We offer over 500 unique circulating analytes, over 100 intracellular analytes, more than 100 kits with more than 30 kits premixed for your convenience. We offer kits covering the broadest range of species: human, non-human primate, mouse, rat, canine, feline, porcine, and equine.

8. What if I cannot find the analyte I need?

First, feel free to contact Technical Support for any questions you may have. In addition, if we do not have your specific biomarker of interest, we offer custom assay services to build MILLIPLEX® MAP kits for your research needs. Contact customassay@emdmillipore.com for a complimentary feasibility assessment. We also offer alternate technologies that may address your needs, such as Single Molecule Counting (SMC™) kits or ELISAs.

9. Can I mix one or more kits together?

Each kit is developed and validated to perform specifically with the reagents included and at the suggested sample dilutions. Combining kits can risk unwanted antibody cross-reactivity, incompatible serum matrices and skewed results. Let us help you determine the feasibility of combining kits. Contact our Technical Support scientists, or our Custom Assay Service Team at customassay@emdmillipore.com.

10. Can I mix reagents from the same kit?

Please only mix reagents if the kit reagents are all from the same lot (lot numbers are on each bottle label inside the kit) and the reagents have been stored under the recommended conditions.

11. Can I use a kit meant for one species to detect samples from a different species?

Our Tips and Tricks brochure contains an appendix with all known kit species cross-reactivity data.  You may also contact our Technical Support team. We also recommend you review the literature.

12. Can a MILLIPLEX® MAP kit be used to detect an analyte in saliva, urine or tears?

Our kits have been validated in the samples types indicated in each kit protocol. Typically, our kits are validated for use with plasma or serum samples, cell culture samples or lysate samples. Some of our kits have been specifically validated for urine or CSF (cerebral spinal fluid). Before trying our kits with a new sample type, please contact Technical Support for recommendations.

13. What is meant by a configurable kit?

Most of our MILLIPLEX® MAP kits are configurable, unless otherwise stated. This mean that you may choose one, or any number of the analytes contained in the kit for your research.

14. How does the price of multiplexing compare to running ELISAs?

Each MILLIPLEX® MAP kit has a base price for the kit with one analyte. Each additional analyte adds incrementally to the price. If you are only looking at one or two analytes, an ELISA or two may suit your budget and needs. However, when you interrogate two or more analytes, multiplexing becomes a real cost-saver. If your goal is to save precious sample volume, then multiplexing is key – most of our MILLIPLEX® MAP kits require only 25 µL of sample (sometimes “neat” and sometimes diluted, please reference each protocol) to quantify all your analytes of interest in one easy assay. You save time by running just one assay with the assurance that our kits provide everything you will need for your experiment. You supply the sample and water.

15. One of the reagent vials in my kit is empty or doesn’t contain enough volume to run the assay. Can you help?

We strive to provide the highest quality assays and service. If you’ve encountered the extremely rare under-filled vial we need to know, and of course we will help. Please contact Technical Support, Customer Service, or your Sales Representative to quickly resolve any issues you have with our kits.

16. I spilled the contents of one of the kit reagent vials. Can you help?

We can certainly help. Please contact Technical Support, Customer Service, or your Sales Representative to quickly resolve any issues you may encounter in using one of our kits.

17. I’m new to multiplexing and am just starting to design my first big experiment and/or write a grant proposal for funding. How can you help?

The earlier you consult with us in your experimental design, the better we can help meet your research needs. We will work with you to design the smartest long-term strategy for your work plan – from biomarker and assay selection, to best practices in sample storage. We will provide quotes upon which you can base your funding requests and operating budget.